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https://www.readbyqxmd.com/read/28218835/selection-of-2-fluoro-modified-aptamers-with-optimized-properties
#1
Deepak Thirunavukarasu, Tingjian Chen, Zhixia Liu, Narupat Hongdilokkul, Floyd E Romesberg
RNA or single-stranded DNA aptamers with 2'-F pyrimidines have been pursued to increase resistance to nucleases, and while it seems likely that these and other modifications, including the modification of purines, could be used to optimize additional properties, this has been much less explored because such aptamers are challenging to discover. Using a thermostable DNA polymerase, SFM4-3, which was previously evolved to accept nucleotides with 2'-modifications, we now report the selection of 2'-F purine aptamers that bind human neutrophil elastase (HNE)...
February 20, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28217561/characterisation-of-the-thermostable-protease-aprx-in-strains-of-pseudomonas-fluorescens-and-impact-on-the-shelf-life-of-dairy-products-preliminary-results
#2
Nadia Andrea Andreani, Lisa Carraro, Luca Fasolato, Stefania Balzan, Rosaria Lucchini, Enrico Novelli, Barbara Cardazzo
Bacterial proteases are involved in food spoilage and shelf-life reduction. Among the bacterial proteases, a predominant role in spoilage of dairy products seems to be played by the thermostable metallo-protease AprX, which is produced by various strains of Pseudomonas fluorescens. Differences in AprX enzyme activity among different strains were highlighted, but the most proteolytic strains were not identified. In this study, the presence of the aprX gene was evaluated in 69 strains isolated from food matrices and 18 reference strains belonging to the P...
September 20, 2016: Italian Journal of Food Safety
https://www.readbyqxmd.com/read/28215563/chitosan-immobilized-novel-peroxidase-from-azadirachta-indica-characterization-and-application
#3
Veda P Pandey, Jyoti Rani, Nivedita Jaiswal, Swati Singh, Manika Awasthi, Ajit K Shasany, Sameeksha Tiwari, Upendra N Dwivedi
In the present paper, a peroxidase was purified from the leaves of a medicinal tree, namely Azadirachta indica, to 45.2 folds with overall recovery of 61%. Based on the subunit size, the purified peroxidase was suggested to be a monomeric structure of size 50kDa and exhibited good thermostability as it was fully stable at 65°C for 1hr and also retained about 73% activity at 70°C till 30min. The substrate affinity was found to be in order of guaiacol > pyrogallol > o-dianisidine. The purified peroxidase was found to be insensitive towards high concentrations of Na(+), Ca(2+), Mg(2+) and Mn(2+)...
February 16, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28215323/marine-enzymes-production-and-applications-for-human-health
#4
T Eswara Rao, M Imchen, R Kumavath
Marine microbial enzymes have wide applications in bioindustries. Selection of microorganisms for enzyme production at the industrial level requires good yield and high production rate. A number of enzymes such as amylase, caseinase, lipase, gelatinase, and DNases have been discovered from microbes isolated from extreme marine environments. Such enzymes are thermostable, tolerant to a varied range of pH and other harsh conditions required in industrial applications. Novelty in their structure and characteristics has shown promising scope to the researchers in academia and industry...
2017: Advances in Food and Nutrition Research
https://www.readbyqxmd.com/read/28214513/molecular-dynamics-simulation-study-of-the-effect-of-glycerol-dialkyl-glycerol-tetraether-hydroxylation-on-membrane-thermostability
#5
Carme Huguet, Susanne Fietz, Antoni Rosell-Melé, Xavier Daura, Lionel Costenaro
Archaeal tetraether membrane lipids span the whole membrane width and present two C40 isoprenoid chains bound by two glycerol groups (or one glycerol and calditol). These lipids confer stability and maintain the membrane fluidity in mesophile to extremophile environments, making them very attractive for biotechnological applications. The isoprenoid lipid composition in archaeal membranes varies with temperature, which has placed these lipids in the focus of paleo-climatological studies for about a decade. Non-hydroxylated isoprenoid archaeal lipids are typically used as paleo-thermometry proxies, but recently identified hydroxylated (OH) derivatives have also been proposed as temperature proxies...
February 15, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28213736/6-phosphofructokinase-and-ribulose-5-phosphate-3-epimerase-in-methylotrophic-bacillus-methanolicus-ribulose-monophosphate-cycle
#6
Simone Balzer Le, Tonje Marita Bjerkan Heggeset, Tone Haugen, Ingemar Nærdal, Trygve Brautaset
D-Ribulose-5-phosphate-3-epimerase (RPE) and 6-phosphofructokinase (PFK) catalyse two reactions in the ribulose monophosphate (RuMP) cycle in Bacillus methanolicus. The B. methanolicus wild-type strain MGA3 possesses two putative rpe and pfk genes encoded on plasmid pBM19 (rpe1-MGA3 and pfk1-MGA3) and on the chromosome (rpe2-MGA3 and pfk2-MGA3). The wild-type strain PB1 also encodes putative rpe and pfk genes on plasmid pBM20 (rpe1-PB1 and pfk1-PB1*); however, it only harbours a chromosomal pfk gene (pfk2-PB1)...
February 17, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28213141/thermostability-of-photosystem-i-trimers-and-monomers-from-the-cyanobacterium-thermosynechococcus-elongatus
#7
Vladimir V Shubin, Irina V Terekhova, Yulia V Bolychevtseva, Eithar El-Mohsnawy, Matthias Rögner, Werner Mäntele, Marta J Kopczak, Enela Džafić
The performance of solar energy conversion into alternative energy sources in artificial systems highly depends on the thermostability of photosystem I (PSI) complexes Terasaki et al. (2007), Iwuchukwu et al. (2010), Kothe et al. (2013) . To assess the thermostability of PSI complexes from the thermophilic cyanobacterium Thermosynechococcus elongatus heating induced perturbations on the level of secondary structure of the proteins were studied. Changes were monitored by Fourier transform infrared (FT-IR) spectra in the mid-IR region upon slow heating (1°C per minute) of samples in D2O phosphate buffer (pD 7...
February 6, 2017: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/28211904/structural-and-functional-characterization-of-deep-sea-thermophilic-bacteriophage-gve2-hnh-endonuclease
#8
Likui Zhang, Dandan Xu, Yanchao Huang, Xinyuan Zhu, Mianwen Rui, Ting Wan, Xin Zheng, Yulong Shen, Xiangdong Chen, Kesen Ma, Yong Gong
HNH endonucleases in bacteriophages play a variety of roles in the phage lifecycle as key components of phage DNA packaging machines. The deep-sea thermophilic bacteriophage Geobacillus virus E2 (GVE2) encodes an HNH endonuclease (GVE2 HNHE). Here, the crystal structure of GVE2 HNHE is reported. This is the first structural study of a thermostable HNH endonuclease from a thermophilic bacteriophage. Structural comparison reveals that GVE2 HNHE possesses a typical ββα-metal fold and Zn-finger motif similar to those of HNH endonucleases from other bacteriophages, apart from containing an extra α-helix, suggesting conservation of these enzymes among bacteriophages...
February 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28208415/thermostability-analysis-of-line-tension-associated-nucleation-at-a-gas-liquid-interface
#9
Sanat Kumar Singha, Prasanta Kumar Das, Biswajit Maiti
The influence of line tension on the thermostability of a droplet nucleated from an oversaturated vapor at the interface of the vapor and another immiscible liquid is investigated. Along with the condition of mechanical equilibrium, the notion of extremization of the reversible work of formation is considered to obtain the critical parameters related to heterogeneous nucleation. From the energetic formulation, the critical reversible work of formation is found to be greater than that of homogeneous nucleation for high value of the positive line tension...
January 2017: Physical Review. E
https://www.readbyqxmd.com/read/28207231/7-approaches-to-elimination-of-the-inherent-systematic-errors-in-determination-of-electrophoretic-mobility-by-capillary-electrophoresis
#10
Paweł Mateusz Nowak, Michał Woźniakiewicz, Pawel Koscielniak
Electrophoretic mobility is a basic parameter describing the electromigration of an ionized particle, which is used in many fields of analytical and physicochemical science. Its determination by capillary electrophoresis (CE) using a routine method is intrinsically affected by the generation of Joule heating, entailing a drop in viscosity and possible alteration of the degree of ionization, and also by other commonly overlooked effects: axial electric field distortion and voltage ramping. The aim of this work was to provide the first theoretical overview and experimental comparison of all accessible methods that could be used to prevent these sources of inaccuracy...
February 16, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28193403/thermal-inactivation-kinetics-of-%C3%AE-galactosidase-during-bread-baking
#11
Lu Zhang, Xiao Dong Chen, Remko M Boom, Maarten A I Schutyser
In this study, β-galactosidase was utilized as a model enzyme to investigate the mechanism of enzyme inactivation during bread baking. Thermal inactivation of β-galactosidase was investigated in a wheat flour/water system at varying temperature-moisture content combinations, and in bread during baking at 175 or 205°C. In the wheat flour/water system, the thermostability of β-galactosidase increased with decreased moisture content, and a kinetic model was accurately fitted to the corresponding inactivation data (R(2)=0...
June 15, 2017: Food Chemistry
https://www.readbyqxmd.com/read/28193333/heterologous-expression-of-%C3%AE-glutamyl-transpeptidase-from-bacillus-atrophaeus-gs-16-and-its-application-in-the-synthesis-of-%C3%AE-d-glutamyl-l-tryptophan-a-known-immunomodulatory-peptide
#12
Meenu Saini, Shruti Bindal, Rani Gupta
Gamma-glutamyl transpeptidase from a mesophilic bacterium Bacillus atrophaeus GS-16 (BaGGT) was expressed heterologously in E. coli using pET-51b vector. Maximum production of BaGGT was obtained at 16°C after 16h of IPTG induction and the protein, in its native conformation, was active as a heterooctamer which was composed of four heterodimeric units combined together. One heterodimeric unit constituted two subunits with molecular masses of 45kDa and 21kDa, respectively. The recombinant enzyme was purified by one step His-tag affinity purification protocol with a specific activity of 90U/mg and 5...
April 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/28193329/characterization-and-crystal-structure-of-a-thermostable-glycoside-hydrolase-family-45-1-4-%C3%AE-endoglucanase-from-thielavia-terrestris
#13
Jian Gao, Jian-Wen Huang, Qian Li, Weidong Liu, Tzu-Ping Ko, Yingying Zheng, Xiansha Xiao, Chih-Jung Kuo, Chun-Chi Chen, Rey-Ting Guo
1,4-β-Endoglucanase is one of the most important biocatalysts in modern industries. Here, a glycoside hydrolase (GH) family 45 endoglucanase from thermophilic fungus Theilavia terrestris (TtCel45A) was expressed in Pichia pastoris. The recombinant protein shows optimal activity at 60°C, pH 4-5. The enzyme exhibits extraordinary thermostability that more than 80% activity was detected after heating at 80°C for 2.5h. The high resolution crystal structures of apo-form enzyme and that in complex with cellobiose and cellotetraose were solved to 1...
April 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/28193326/properties-of-a-novel-thermostable-glucose-isomerase-mined-from-thermus-oshimai-and-its-application-to-preparation-of-high-fructose-corn-syrup
#14
Dong-Xu Jia, Lin Zhou, Yu-Guo Zheng
Glucose isomerase (GI) is used in vitro to convert d-glucose to d-fructose, which is capable of commercial producing high fructose corn syrup (HFCS). To manufacture HFCS at elevated temperature and reduce the cost of enriching syrups, novel refractory GIs from Thermoanaerobacterium xylanolyticum (TxGI), Thermus oshimai (ToGI), Geobacillus thermocatenulatus (GtGI) and Thermoanaerobacter siderophilus (TsGI) were screened via genome mining approach. The enzymatic characteristics research showed that ToGI had higher catalytic efficiency and superior thermostability toward d-glucose among the screened GIs...
April 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/28193276/thermus-thermophilus-as-source-of-thermozymes-for-biotechnological-applications-homologous-expression-and-biochemical-characterization-of-an-%C3%AE-galactosidase
#15
Martina Aulitto, Salvatore Fusco, Gabriella Fiorentino, Danila Limauro, Emilia Pedone, Simonetta Bartolucci, Patrizia Contursi
BACKGROUND: The genus Thermus, which has been considered for a long time as a fruitful source of biotechnological relevant enzymes, has emerged more recently as suitable host to overproduce thermozymes. Among these, α-galactosidases are widely used in several industrial bioprocesses that require high working temperatures and for which thermostable variants offer considerable advantages over their thermolabile counterparts. RESULTS: Thermus thermophilus HB27 strain was used for the homologous expression of the TTP0072 gene encoding for an α-galactosidase (TtGalA)...
February 13, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28192949/one-pot-synthesis-of-fluorescent-silicon-nanoparticles-for-sensitive-and-selective-determination-of-2-4-6-trinitrophenol-in-aqueous-solution
#16
Yangxia Han, Yonglei Chen, Jie Feng, Juanjuan Liu, Sudai Ma, Xingguo Chen
Because 2,4,6-trinitrophenol (TNP) and its analogues such as 2,4,6-trinitrotoluene (TNT) possess similar chemical structures and properties, the reliable and accurate detection of TNP from its analogues still remains a challenging task. In the present work, a selective and sensitive method based on the water-soluble silicon nanoparticles (SiNPs) for the determination of TNP was established. The SiNPs with good thermostability and excellent antiphotobleaching capability were prepared via a simple one-pot method...
February 21, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28188780/structural-insights-into-the-cold-adaptation-of-the-photosynthetic-pigment-protein-c-phycocyanin-from-an-arctic-cyanobacterium
#17
Hai-Nan Su, Qian-Min Wang, Chun-Yang Li, Kang Li, Wei Luo, Bo Chen, Xi-Ying Zhang, Qi-Long Qin, Bai-Cheng Zhou, Xiu-Lan Chen, Yu-Zhong Zhang, Bin-Bin Xie
The cold adaptation mechanism of phycobiliproteins, the major photosynthetic pigment-proteins in cyanobacteria and red algae, has rarely been studied. Here we reported the biochemical, structural, and molecular dynamics simulation study of the C-phycocyanin from Arctic cyanobacterial strain Pseudanabaena sp. LW0831. We characterized the phycobilisome components of LW0831 and obtained their gene sequences. Compared to the mesophilic counterpart from Arthrospira platensis (Ar-C-PC), LW0831 C-phycocyanin (Ps-C-PC) has a decreased thermostability (∆Tm of -16°C), one of the typical features of cold-adapted enzymes...
February 7, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28187141/role-of-n-linked-glycosylation-in-the-enzymatic-properties-of-a-thermophilic-gh-10-xylanase-from-aspergillus-fumigatus-expressed-in-pichia-pastoris
#18
Xiaoyu Chang, Bo Xu, Yingguo Bai, Huiying Luo, Rui Ma, Pengjun Shi, Bin Yao
N-Glycosylation is a posttranslational modification commonly occurred in fungi and plays roles in a variety of enzyme functions. In this study, a xylanase (Af-XYNA) of glycoside hydrolase (GH) family 10 from Aspergillus fumigatus harboring three potential N-glycosylation sites (N87, N124 and N335) was heterologously produced in Pichia pastoris. The N-glycosylated Af-XYNA (WT) exhibited favorable temperature and pH optima (75°C and pH 5.0) and good thermostability (maintaining stable at 60°C). To reveal the role of N-glycosylation on Af-XYNA, the enzyme was deglycosylated by endo-β-N-acetylglucosaminidase H (DE) or modified by site-directed mutagenesis at N124 (N124T)...
2017: PloS One
https://www.readbyqxmd.com/read/28185055/improving-pullulanase-catalysis-via-reversible-immobilization-on-modified-fe3o4-polydopamine-nanoparticles
#19
Jianfeng Wang, Zhongmei Liu, Zhemin Zhou
To improve the catalysis of pullulanase from Anoxybacillus sp.WB42, Fe3O4@polydopamine nanoparticles (Fe3O4@PDA) were prepared and modified with functional groups for immobilization of pullulanases via covalent binding or ionic adsorption. Immobilized pullulanases had lower thermal stability than that of free pullulanase, whereas their catalysis depended on the surface characteristics of nanoparticles. As for covalent immobilization of pullulanases onto Fe3O4@PDA derivatives, the spacer grafted onto Fe3O4@PDA made the catalytic efficiency of pullulanase increase up to the equivalence of free enzyme but dramatically reduced the pullulanase thermostability...
February 9, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28184988/cloning-evaluation-and-high-level-expression-of-a-thermo-alkaline-pectate-lyase-from-alkaliphilic-bacillus-clausii-with-potential-in-ramie-degumming
#20
Cheng Zhou, Yanfen Xue, Yanhe Ma
Alkaline pectate lyases (Pels) have potential application in bioscouring of the textile industry. In this study, a thermo-alkaline Pel (BacPelA) gene from an alkaliphilic Bacillus clausii strain was cloned and overexpressed in Escherichia coli. The mature BacPelA exhibited maximum activity at pH 10.5 and 70 °C and showed high cleavage capability on methylated pectins. BacPelA showed the highest specific activity of 936.2 U mg(-1) on ≥85% methylated pectin and 675.5 U mg(-1) on standard substrate polygalacturonic acid (PGA) upon evaluation of the absorbance at 235 nm (A235)...
February 9, 2017: Applied Microbiology and Biotechnology
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